Instrument: Illumina HiSeq 4000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: The pre-cultured cells of BY4741 and BY4741(yrr1Δ) in SD medium were transferred into fresh SD or SD added 5mM vanillin with an initial OD600 of 0.2. The cells were harvested during the log phase (OD600≈1.6-2.0) and quick frozen using liquid nitrogen. UNIQ-10 Trizol RNA Purification Kit (Sangon Biotech, China) was used to extract total RNA. After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT) are used to isolate mRNA(for eukaryotes) or by removing rRNAs from the total RNA (for prokaryotes). Mixed with the fragmentation buffer, the mRNA is fragmented into short fragments. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates.After that, the short fragments are connected with adapters. After agarose gel electrophoresis, the suitable fragments are selected for the PCR amplification as templates.